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1.
BMC Infect Dis ; 21(1): 439, 2021 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-33985447

RESUMO

BACKGROUND: Genetic diversity in Plasmodium falciparum populations can be used to describe the resilience and spatial distribution of the parasite in the midst of intensified intervention efforts. This study used microsatellite analysis to evaluate the genetic diversity and population dynamics of P. falciparum parasites circulating in three ecological zones of Ghana. METHODS: A total of 1168 afebrile children aged between 3 to 13 years were recruited from five (5) Primary schools in 3 different ecological zones (Sahel (Tamale and Kumbungu), Forest (Konongo) and Coastal (Ada and Dodowa)) of Ghana. Asymptomatic malaria parasite carriage was determined using microscopy and PCR, whilst fragment analysis of 6 microsatellite loci was used to determine the diversity and population structure of P. falciparum parasites. RESULTS: Out of the 1168 samples examined, 16.1 and 39.5% tested positive for P. falciparum by microscopy and nested PCR respectively. The genetic diversity of parasites in the 3 ecological zones was generally high, with an average heterozygosity (He) of 0.804, 0.787 and 0.608 the rainy (peak) season for the Sahel, Forest and Coastal zones respectively. The mean He for the dry (off-peak) season were 0.562, 0.693 and 0.610 for the Sahel, Forest and Coastal zones respectively. Parasites from the Forest zone were more closely related to those from the Sahel than from the Coastal zone, despite the Coastal zone being closer in physical distance to the Forest zone. The fixation indexes among study sites ranged from 0.049 to 0.112 during the rainy season and 0.112 to 0.348 during the dry season. CONCLUSION: A large asymptomatic parasite reservoir was found in the school children during both rainy and dry seasons, especially those in the Forest and Sahel savannah zones where parasites were also found to be related compared to those from the Coastal zone. Further studies are recommended to understand why despite the roll out of several malaria interventions in Ghana, high transmission still persist.


Assuntos
Portador Sadio/parasitologia , Malária Falciparum/parasitologia , Plasmodium falciparum/genética , Adolescente , Portador Sadio/epidemiologia , Criança , Pré-Escolar , DNA de Protozoário/genética , Feminino , Variação Genética , Genética Populacional , Gana/epidemiologia , Humanos , Malária Falciparum/epidemiologia , Masculino , Repetições de Microssatélites/genética , Plasmodium falciparum/citologia , Plasmodium falciparum/isolamento & purificação , Estações do Ano
2.
Ann Parasitol ; 67(1): 39-44, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34010549

RESUMO

Leishmaniosis is one of the most important vector borne diseases. Among different forms of the disease, cutaneous leishmaniosis (CL) is the most common. Determining the method of definitive diagnosis for the disease has been the aim of various studies. Therefore this study afforded an opportunity to investigate this subject. To diagnose CL in 150 suspected patients referred to Mehran and Dehloran health centers during June 2018 to November 2019, two polymerase chain reaction (PCR) methods were performed and compared with the in vitro culture and microscopic evaluation of stained slides. The smears were stained with Giemsa for microscopy and cultured in Novy-Nicolle-McNeal (NNN) blood agar for promastigote growth. For semi-nested PCR and PCR-RFLP, the tissue and serosity from the lesions were used for DNA extraction. The semi-nested PCR technique using minicircle kDNA gene showed the highest positivity rates among all diagnostic assays with 114/150 (76%) of the samples and was used as reference standard, followed by the PCR-RFLP test using ITS1 gene with 112/150, (74.7%) positivity rates, microscopy with 101/150 (67.3%) and then culture 72/150 (48%). microscopy and culture methods together improved overall positivity rates to 68.7% (103/150). The all positive samples using molecular technique were identified as Leishmania major. The highest sensitivity (98.3%), specificity (100%), accuracy (98.8%), negative predictive value (94.7%) and κ coefficient (0.96=almost perfect) was observed by comparing PCR-RFLP and semi-nested PCR. kDNA-semi-nested PCR and ITS1-PCR-RFLP presented an interesting alternative to conventional methods for the identification of CL and improved its diagnostic value significantly in suspected patients with negative direct smears.


Assuntos
Leishmania major , Leishmaniose Cutânea , DNA de Cinetoplasto , DNA de Protozoário , Humanos , Leishmaniose Cutânea/diagnóstico , Reação em Cadeia da Polimerase , Projetos de Pesquisa
3.
BMC Infect Dis ; 21(1): 464, 2021 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-34020601

RESUMO

BACKGROUND: Leishmaniasis is one of the most neglected tropical diseases in the world and remains endemic in some underdeveloped regions, including western China. The phylogeny and classification of Chinese Leishmania has not been completely clarified to date, especially within the Leishmania (L.) donovani complex, although phylogenetic analyses based on a series of gene markers have been performed. More analytic methods and data are still needed. Random amplified polymorphic DNA (RAPD) technology can sensitively identify slight intraspecific differences, and it is a powerful tool to seek species-specific markers. This work attempted to identify Chinese Leishmania isolates from diverse geographic regions at the genomic level. Meanwhile, specific markers of the L. donovani complex were also developed by RAPD. METHODS: RAPD was applied to 14 Chinese Leishmania isolates from diverse geographic regions and 3 WHO reference strains. The polymorphic sites of amplification were transformed into a data matrix, based on which genetic similarity was calculated, and a UPGMA dendrogram was constructed to analyse the genetic diversity of these Leishmania isolates. Meanwhile, the specific amplification loci of the L. donovani complex were TA-cloned, sequenced and converted into sequence characterized amplified region (SCAR) markers, which were validated preliminarily in 17 available Leishmania strains in this study and analysed by bioinformatics. RESULTS: The cluster analyses showed that the three Leishmania sp. isolates SC10H2, SD and GL clustered together and apart from others, the strains of the L. donovani complex clearly divided into two clades, and the three isolates Cy, WenChuan and 801 formed a subclade. Three specific SCAR markers of the L. donovani complex, i.e., 1-AD17, 2-A816 and 3-O13, were successfully obtained and validated on 17 available Leishmania strains in this study. Through bioinformatic analyses, Marker 1-AD17 may have more specificity for PCR detection of VL, and Marker 3-O13 has the potential to encode a protein. CONCLUSIONS: The RAPD results verified that the undescribed Leishmania species causing visceral leishmaniasis (VL) in China was a unique clade distinguished from L. donovani and revealed that there was genetic differentiation among Chinese L. donovani. The identification of L. donovani-specific markers may help to provide a foundation for future research attempting to develop new specific diagnostic markers of VL and identify specific gene functions.


Assuntos
Variação Genética , Leishmania donovani/classificação , Leishmania donovani/genética , Leishmaniose Visceral/epidemiologia , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Animais , Sequência de Bases , China/epidemiologia , Análise por Conglomerados , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Marcadores Genéticos , Humanos , Leishmania donovani/isolamento & purificação , Leishmaniose Visceral/parasitologia , Filogenia , Reação em Cadeia da Polimerase , Especificidade da Espécie
4.
Rev Bras Parasitol Vet ; 30(2): e000621, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33978118

RESUMO

Molecular methods such as Copro-PCR stand out in the diagnosis of T. gondii, because they are highly sensitive and specific, and can distinguish T. gondii from other morphologically similar coccids. The purpose was the detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction in 149 fecal samples from stray and domiciled cats, using three distinct markers (B5-B6, 18S and 529bp RE). Oocysts of T. gondii/H. hammondi were detected in 15.4% by parasitology fecal tests (PFT), and 4% of these oocysts were positively identified as T. gondii by Copro-PCR. The presence of T. gondii genetic material was detected in 16.1%, but 12% of the samples that tested positive by Copro-PCR were negative in PFT. Samples with discordant results were subjected to a new Copro-PCR with 18S marker and a 529, and of the 17 samples, 9 contained T. gondii genetic material. A comparison of the PFT and the molecular methods showed the latter was more sensitive, since it detected 22.1% while the PFT detected 15.4%. Demonstrating the high sensitivity and specificity of the Copro-PCR, particularly with the association of primers (k=0.809), but also confirms the importance of using molecular techniques in laboratories, since Copro-PCR was able to detect samples considered negative by PFT.


Assuntos
Doenças do Gato , Toxoplasma , Toxoplasmose Animal , Animais , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Gatos , DNA de Protozoário , Fezes , Oocistos , Reação em Cadeia da Polimerase/veterinária , Toxoplasma/genética , Toxoplasmose Animal/diagnóstico , Toxoplasmose Animal/epidemiologia
5.
Exp Parasitol ; 225: 108113, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33992605

RESUMO

Cryptosporidiosis remains the leading protozoan induced cause of diarrhoea-associated mortality worldwide. Cryptosporidium hominis, the anthroponotically transmitted species within the Cryptosporidium genus, contributes significantly to the global burden of infection, accounting for the majority of clinical cases in many countries. This study applied high resolution melting analysis, a post-real-time PCR application, to the differentiation of six globally prevalent C. hominisgp60-subtypes. This novel method targeted three microsatellite, tandem repeat containing genetic markers, gp60, the genetic marker upon which current Cryptosporidium subtype nomenclature is based, MSB, and MSE, by which to differentiate between C. hominis isolates. This multi-locus approach successfully differentiated between all six C. hominisgp60-subtypes studied, some of which, such as IbA10G2, are known to exhibit global ubiquity. Thus, this method has the potential to be universally employed as a sensitive, cost effective and highly reproducible means to rapidly differentiate between C. hominisgp60-subtypes. Such a method would be of particular utility in epidemiological studies and outbreak scenarios, providing cost effective, clinically accessible alternative to DNA sequencing. The success of this preliminary study also supports further analysis of an expanded C. hominisgp60-subtype range and the potential expansion of the multi-locus panel in order to improve the discriminatory power of this approach.


Assuntos
Cryptosporidium/genética , Parasitologia/métodos , Criptosporidiose/parasitologia , Cryptosporidium/classificação , Cryptosporidium/isolamento & purificação , DNA de Protozoário/genética , Fezes/parasitologia , Marcadores Genéticos , Genótipo , Humanos , Tipagem de Sequências Multilocus , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA
6.
Artigo em Inglês | MEDLINE | ID: mdl-33909846

RESUMO

Blastocystis sp. is an enteric protist commonly found in human fecal samples. In Brazil, few studies have been developed, but none of them has explored the presence of Blastocystis in patients with diabetes mellitus. We evaluated the occurrence and molecular identification of Blastocystis sp. among patients with diabetes mellitus in the Midwest region, Goias State, Brazil. Genomic DNA was obtained from 175 fecal samples (99 from the diabetic group and 76 from the control group). PCR was performed using pan-Blastocystis primers from the SSU-rDNA gene. Microscopic examination revealed positivity of 12.1% and 7.9% for Blastocystis in diabetics and in controls, respectively. Amplification of Blastocystis DNA was observed in 34.4% (34 of 99) and 30.3% (23 of 76) from the diabetic and control groups, respectively. Phylogenetic analyses and BLAST searches revealed six subtypes among Blastocystis isolates in the diabetic group, represented by ST1 (38.2%), ST2 (11.8%), ST3 (35.3%), ST6 (2.9%), ST7 (2.9%) and ST8 (8.8%). In the control group, ST1 (21.8%), ST2 (21.8%), ST3 (43.5%), ST6 (4.4%) and ST8 (8.7%) were identified. This study is the first report regarding the occurrence and subtypes distribution of Blastocystis in patients with diabetes mellitus in Brazil. The results reinforce the potential risk of Blastocystis infection in patients with diabetes, in addition, it contributes to the understanding of the genetic diversity of this enigmatic organism.


Assuntos
Infecções por Blastocystis , Blastocystis , Diabetes Mellitus , Blastocystis/genética , Infecções por Blastocystis/epidemiologia , Brasil/epidemiologia , DNA de Protozoário/genética , Diabetes Mellitus/epidemiologia , Fezes , Variação Genética , Humanos , Filogenia
7.
Mem Inst Oswaldo Cruz ; 116: e200572, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33886871

RESUMO

BACKGROUND: The genetic heterogeneity of Leishmania parasites is a major factor responsible for the wide variety of Leishmania-associated manifestations. Consequently, understanding the genetic make-up of Leishmania species using suitable molecular markers is an important component of realising local and regional scale disease risk. The cytochrome b (cytb) is frequently used to type New World Leishmania species. However, its potential to discriminate Leishmania species and variants requires further evaluation. OBJECTIVES: To explore the capacity of cytb gene to identify New World Leishmania species and variants and to develop an approach able to type local Leishmania species and variants. METHODS: We retrieved 360 partial and complete Leishmania cytb gene sequences publicly available in GenBank database to study all single nucleotide polymorphisms (SNPs) across the cytb gene that differentiate New World Leishmania species. This information was used to develop an approach based upon the polymorphisms found in a DNA segment of 948bp. We also compared the typing results found with this technique with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) profiling obtained using HSP70 gene as target. One hundred Panamanian isolates were used to both typed Leishmania species and assess local genetic variability. FINDINGS: We found complete agreement between our cytb approach and the PCR-RFLP profiling method based on HSP70 for Leishmania species identification. Ninety-two isolates were identified as L. panamensis, although other Viannia species were found circulating at a lower frequency. Three L. panamensis haplotypes were identified in Panamanian provinces. We also provide an initial report of L. guyanensis haplotypes circulating in Panama. MAIN CONCLUSIONS: Cytb gene sequence encompasses key main SNPs that aid to identify Leishmania species. The cytb approach developed with this information was able to identify and assess genetic variability of local Leishmania species found in this study.


Assuntos
Leishmania , Leishmaniose Cutânea , Citocromos b/genética , DNA de Protozoário/genética , Humanos , Leishmania/genética , Panamá , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição
8.
Parasitol Res ; 120(5): 1837-1844, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33649965

RESUMO

Cryptosporidium is an important intestinal protozoan parasite that causes diarrhoea in humans and animals. To rapidly and specifically detect Cryptosporidium spp., we designed a pair of primers based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. to be used in a new nanoparticle-assisted PCR (nano-PCR) assay. The minimum detectable concentration (1.02 pg) of this nano-PCR was 10 times more sensitive than conventional PCR using the same primer pair. The DNA samples of C. parvum, C. baileyi, C. xiaoi, C. ryanae, and C. andersoni were successfully detected by the nano-PCR. No amplifications were evident with DNA samples of some common intestinal pathogens, including Eimeria tenella, Blastocystis sp., Giardia lamblia, Enterocytozoon bieneusi, and Balantidium coli. To validate the clinical usefulness of the novel nano-PCR, a total of 40 faecal samples from goats, camels, calves, and chickens were examined. The positive rate of Cryptosporidium spp. was 27.5% (11/40), which was consistent with that of an established nested PCR. These results indicate that the novel nano-PCR assay enables the rapid, specific, and accurate detection of Cryptosporidium infection in animals. The findings provide a technical basis for the clinical diagnosis, prevention, and control of cryptosporidiosis.


Assuntos
Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , Nanopartículas , Reação em Cadeia da Polimerase/métodos , Animais , Camelus , Bovinos , Galinhas , Criptosporidiose/parasitologia , Cryptosporidium/genética , Cryptosporidium parvum/genética , DNA de Protozoário , Fezes/parasitologia , Cabras , Análise de Sequência de DNA
9.
Parasitol Res ; 120(5): 1873-1882, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33712930

RESUMO

The early containment of trypanosomosis depends on early, sensitive, and accurate diagnosis in endemic areas with low-intensity infections. The study was planned to develop a simple read out loop-mediated isothermal amplification (LAMP) assay targeting a partial RoTat1.2 VSG gene of Trypanosoma evansi with naked eye visualization of LAMP products by adding SYBR® Green I dye. The visual results were further confirmed with those of agarose gel electrophoresis, restriction enzyme digestion of LAMP products with AluI, and sequencing of the PCR products using LAMP outer primers. The LAMP primers did not show cross reactivity and non-specific reactions with regional common hemoparasitic DNA revealing high specificity of the assay. The threshold sensitivity level of the LAMP assay was determined to be 0.003 fg compared to 0.03 fg RoTat1.2 amplified DNA fragments of T. evansi by PCR assay. Moreover, assessment of 500 blood samples collected from unhealthy domestic animals in field suspected for various hemoparasitic infections was carried out for the presence of T. evansi by microscopy, RoTat1.2 VSG PCR, and LAMP assay. LAMP could detect T. evansi in 36 samples, while PCR and microscopy could detect 33 and 12 samples, respectively. All the samples positive by microscopy and PCR were also confirmed positive by the LAMP assay. The current LAMP assay has appealing point of care characteristics to visually monitor the results, lessen the need of post DNA amplification procedure, and enable this method to be applied as a rapid and sensitive molecular diagnostic tool in under resourced laboratories and field setup.


Assuntos
Antígenos de Protozoários/genética , Técnicas de Diagnóstico Molecular/veterinária , Técnicas de Amplificação de Ácido Nucleico/veterinária , Proteínas de Protozoários/genética , Trypanosoma/genética , Trypanosoma/isolamento & purificação , Animais , Animais Domésticos/parasitologia , Primers do DNA , DNA de Protozoário/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Tripanossomíase/parasitologia
10.
Parasitol Res ; 120(5): 1915-1919, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33740119

RESUMO

This study reports the identification and first molecular characterization of Babesia occultans from naturally infected cows in Iran. Microscopic examination showed pyriform trophozoites, and ring-shaped merozoites (>2.5 µm) in Giemsa-stained blood smears obtained from two symptomatic cows in West-Azarbaijan province, Iran. PCR amplification of the partial 18S rRNA gene including the V4 hypervariable region were carried out on DNA extracted from blood samples. BLAST analyses of the partial 18S rRNA (approximately 400 bp) obtained from two cows revealed the presence of B. occultans and the detected sequences were identical to each other. Comparisons of the partial 18S rRNA sequence of the current isolate with other B. occultans sequences from Tunisia, South Africa, Turkey, Pakistan, and China confirmed the relation of the Iranian isolate to the species B. occultans. Sequence analysis of the obtained B. occultans showed 99.5-100% identity to the previously reported isolates. The sequences of B. occultans had 100% identity to a sequence obtained from ticks in Tunisia. This report is the beginning of a path to further research about B. occultans in vectors and reservoirs throughout Iran.


Assuntos
Babesia/isolamento & purificação , Babesiose/parasitologia , Doenças dos Bovinos/parasitologia , Animais , Babesia/genética , Babesiose/sangue , Bovinos , DNA de Protozoário/genética , Feminino , Irã (Geográfico) , Filogenia , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S , Carrapatos/genética
11.
Acta Trop ; 218: 105905, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33775628

RESUMO

Haemoproteus species (Haemosporida, Haemoproteidae) are cosmopolitan blood parasites, which have been neglected for over 100-years, but attracted attention recently due to reports of severe and even lethal haemoproteosis in birds and vectors. Approximately 150 species of avian Haemoproteus have been described and named, but molecular data suggest that hundreds of independently evolving molecular lineages might occur, indicating the existence of a remarkable undescribed species diversity. It is timely to develop a methodology, which allow the application of available genetic data in taxonomy of haemosporidians on species levels. This study aimed to test a hypothesis suggesting that DNA haplotype networks might aid in targeting genetically distinct, but still undescribed parasites, and might be used to direct taxonomic studies on haemosporidian species levels. Mainly, we tested a prediction that the lineage hTUPHI01, a common Haemoproteus parasite of Turdus philomelos, might be a new species, which is morphologically similar and genetically closely related to the parasites of Haemoproteus minutus group. Blood samples of T. philomelos naturally infected with this parasite lineage were collected and studied using microscopic examination of blood films and PCR-based methods. Haemoproteus asymmetricus n. sp. was found in this bird, described and characterised molecularly using partial cytochrome b (cytb) sequences. The new species shared some features with parasites of the H. minutus group, as was predicted by the DNA haplotype network. Due to the visualisation of closely related lineages as well as the evaluation of their host and geographic distributions, DNA haplotype networks can be recommended as the helpful methodology, able to direct and speed practical work on parasite species taxonomy and pathogen biodiversity. The combined molecular phylogenetic and morphological approaches showed that the well-supported clades in Bayesian phylogenetic trees based on the partial cytb gene sequences contain morphologically remarkably different Haemoproteus parasite species, which however, share some basic biological features. Phylogenetic analysis can be used for prediction of these basic features in still undescribed parasites. This study calls for further fusion of advanced molecular and microscopy approaches for better understanding haemosporidian parasite biology.


Assuntos
Doenças das Aves/parasitologia , Aves/parasitologia , Haemosporida/citologia , Haemosporida/genética , Infecções Protozoárias em Animais/parasitologia , Animais , Teorema de Bayes , Doenças das Aves/sangue , Aves/sangue , Citocromos b/genética , DNA de Protozoário , Testes Diagnósticos de Rotina , Gametogênese , Genes de Protozoários , Genoma de Protozoário , Haemosporida/classificação , Haplótipos , Tipagem Molecular , Filogenia , Reação em Cadeia da Polimerase
12.
Acta Trop ; 218: 105907, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33789154

RESUMO

PURPOSE: Toxoplasma gondii is an intracellular parasite that can affect all vertebrae and is the causative agent of toxoplasmosis. At present, the United States CDC (Centers for Disease Control and Prevention) recognizes this infection as a neglected disease. Toxoplasma gondii infection profiles exhibit differences because of the different regional and climatic responses to these parasites in Turkey, and these protozoan infections are notably common in this country. In this study, we attempted to obtain the whole-genome sequence of T. gondii using next-generation sequencing technology. METHODS: Toxoplasma gondii isolates were isolated from an infant with congenital toxoplasmosis by Ekmen et al. (1974) in Ankara, Turkey. Whole-genome sequencing (WGS) was performed using the Illumina HiSeq 2500 and HiSeq SBS Kit v2. A T. gondii library was created on this device in the initial stage. After the completion of the library phase, sequence analysis was begun with a next-generation sequencing device. The resulting fragments were combined using paired-end (PE) reading and converted into a single DNA fragment. Bioinformatic analysis was performed using the Geneious 2.1. RESULTS: In our study, WGS was successfully performed on T. gondii. The T. gondii whole-genome sequence has a coverage value of 50x, a size of 61,5763 Mb and a GC ratio of 52.6%. Data from this sequence were submitted to the National Center for Biotechnology Information (NCBI) GenBank (www.ncbi.nlm.nih.gov) database under the name Toxoplasma gondii TR01 (TG_TR01). The accession number of the genome obtained in this study is WOEV00000000.1. The biological sample access number is SAMN13338796. The genome of the T. gondii strain obtained in this study was compared with the reference genome, and 8312 CDSs (coding sequences), 183 tRNAs, 294 rRNAs and 8789 genes were identified. Among the 8312 CDSs, 4284 encoded hypothetical proteins (hypothetical protein CDSs/proteins of unknown function). The entire genome sequence of T gondii TR01 was compared with that of Toxoplasma gondii ME49. The results of this comparison demonstrate that the analyzed genome was 99,98% similar to the reference genome. The accession numbers of 14 chromosomes belonging to the genome sequences of T. gondii TR01 (TG_TR01) are CM019722.1, CM019723.1, CM019724.1, CM019725.1, CM019726.1, CM019727.1, CM019728.1, CM019729.1, CM019730.1, CM019731.1, CM019732.1, CM019733.1, CM019734.1, and CM019735.1. CONCLUSION: In this study, a whole-genome sequences of T. gondii was conducted for the first time in Turkey. The analyzed strain was named T. gondii TR01. The data obtained from this study may contribute to a better understanding of T. gondii. T. gondii is an important pathogen with an unusual population structure. Although T. gondii is highly zoonotic and has a complicated life cycle, some strains of this parasite have exhibited high genetic sequence similarity, and our study supports this knowlegde. The characterization of this strain may be very useful for the scientific community of our country and may help to establish a foundation for further research investigating the genome of T. gondii.


Assuntos
Toxoplasma/genética , Toxoplasmose Congênita/parasitologia , Animais , Biologia Computacional , DNA de Protozoário , Biblioteca Gênica , Genoma de Protozoário , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Camundongos , Análise de Sequência de DNA , Turquia , Sequenciamento Completo do Genoma
13.
Turkiye Parazitol Derg ; 45(1): 11-16, 2021 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-33685062

RESUMO

Objective: This study aimed to determine the prevalence of canine visceral leishmaniasis by molecular and serological techniques among owned dogs brought to veterinary clinics in the Mugla region of Turkey. Methods: Blood samples were collected from a total of 131 dogs of different breeds and gender that were brought to veterinary clinics between October 2017 and November 2018 in the Mugla region. These blood samples were analysed using immunofluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Results: According to the IFAT results, 49 out of 131 dogs (37.4%) were found to have anti-Leishmania antibodies at a titer of ≥1/64, which was considered as seropositive. On the other hand, PCR results obtained in this study showed that 9 out of 131 dogs (6.87%) were Leishmania spp. positive by RV1/RV2 PCR. Conclusion: The results suggest that there is a need to focus on developing measures to control the disease, fight the vector Phlebotom and initiate disease control programmes for dogs in the Mugla region.


Assuntos
Doenças do Cão/epidemiologia , Leishmania/isolamento & purificação , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antiprotozoários/sangue , DNA de Protozoário/genética , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Leishmania/genética , Leishmania/imunologia , Leishmaniose Visceral/sangue , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Prevalência , Turquia/epidemiologia
14.
Zootaxa ; 4942(2): zootaxa.4942.2.9, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33757071

RESUMO

Anteholosticha sigmoidea (Foissner, 1982) Berger, 2003 was isolated from a wet soil sample collected on King George Island, Antarctica. Morphological observations and molecular phylogenetic analyses based on the gene sequences of small subunit ribosomal RNA (18S rRNA) were used to identify the species. Anteholosticha sigmoidea can be divided into two groups: group I (three populations described by Foissner 1982) and group II (described by Foissner 1984) based on the morphological differences. Group I differs from group II by the length of the midventral complex (65.1% vs. 52.5% of the cell length), the number of adoral membranelles (25-28 vs. 16-24), and the number of dorsal bristles in kinety 1 (16 bristles vs. nine bristles). Group I differs from the Antarctica population by the absence/presence of the collecting canals of the contractile vacuole and the number of macronuclear nodules (6-12 vs. 13-19). Group II differs from the Antarctica population by the number of macronuclear nodules (five to nine vs. 13-19); the arrangement of cortical granules (forming longitudinal rows vs. irregularly distributed); the length of the midventral complex (64.7% vs. 53.8% of cell length). In the phylogenetic analyses, A. sigmoidea was not nested with any species, and the gene tree indicated polyphyly of the genus Anteholosticha.


Assuntos
Cilióforos , Animais , Regiões Antárticas , Cilióforos/genética , DNA de Protozoário , Filogenia , RNA Ribossômico 18S/genética
15.
BMC Infect Dis ; 21(1): 259, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33711940

RESUMO

BACKGROUND: Plasmodium cynomolgi is a simian malaria parasite that has been reported as a naturally acquired human infection. The present study aims to systematically review reports on naturally acquired P. cynomolgi in humans, mosquitoes, and macaques to provide relevant data for pre-emptive surveillance and preparation in the event of an outbreak of zoonotic malaria in Southeast Asia. METHODS: The protocol of the systematic review was registered at PROSPERO with approval ID CRD42020203046. Three databases (Web of Science, Scopus, and MEDLINE) were searched for studies reporting the prevalence of P. cynomolgi infections in Southeast Asian countries between 1946 and 2020. The pooled prevalence or pooled proportion of P. cynomolgi parasitemia in humans, mosquitoes, and macaques was estimated using a random-effects model. Differences in the clinical characteristics of P. cynomolgi infections were also estimated using a random-effects model and presented as pooled odds ratios (ORs) or mean differences (MDs) with 95% confidence intervals (CIs). RESULTS: Thirteen studies reporting on the prevalence of naturally acquired P. cynomolgi in humans (3 studies, 21 cases), mosquitoes (3 studies, 28 cases), and macaques (7 studies, 334 cases) were included. The results demonstrated that the pooled proportion of naturally acquired P. cynomolgi in humans was 1% (95% CI, 0.1%, I2, 0%), while the pooled proportion of P. cynomolgi infecting mosquitoes was 18% (95% CI, 10-26%, I2, 32.7%). The pooled prevalence of naturally acquired P. cynomolgi in macaques was 47% (95% CI, 27-67%, I2, 98.3%). Most of the cases of naturally acquired P. cynomolgi in humans were reported in Cambodia (62%) and Malaysia (38%), while cases of P. cynomolgi in macaques were reported in Malaysia (35.4%), Singapore (23.2%), Indonesia (17.3%), Philippines (8.5%), Laos (7.93%), and Cambodia (7.65%). Cases of P. cynomolgi in mosquitoes were reported in Vietnam (76.9%) and Malaysia (23.1%). CONCLUSIONS: This study demonstrated the occurrence of naturally acquired P. cynomolgi infection in humans, mosquitoes, and macaques. Further studies of P. cynomolgi in asymptomatic human cases in areas where vectors and natural hosts are endemic are extensively needed if human infections with P. cynomolgi do become public health problems.


Assuntos
Culicidae/parasitologia , Macaca/parasitologia , Malária/diagnóstico , Plasmodium cynomolgi/isolamento & purificação , Animais , Ásia Sudeste/epidemiologia , DNA de Protozoário/metabolismo , Humanos , Malária/epidemiologia , Razão de Chances , Plasmodium cynomolgi/genética , Prevalência
16.
Rev Bras Parasitol Vet ; 30(1): e022020, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33729316

RESUMO

Leishmaniasis is a zoonotic disease caused by over 20 species of protozoan parasites of the genus Leishmania. Infection is commonly spread by sandflies and produces a wide spectrum of clinical signs and symptoms. Therefore, from an epidemiological and therapeutic standpoint, it is important to detect and differentiate Leishmania spp. The objective of this study was to combinate in silico and in vitro strategies to evaluate the analytical specificity of primers previously described in the literature. According to electronic PCR (e-PCR) analysis, 23 out of 141 pairs of primers selected through literature search matched their previously reported analytical specificity. In vitro evaluation of nine of these primer pairs by quantitative PCR (qPCR) confirmed the analytical specificity of five of them at the level of Leishmania spp., L. mexicana complex or Leishmania and Viannia subgenera. Based on these findings, the combination of e-PCR and qPCR is suggested to be a valuable approach to maximize the specificity of new primer pairs for the laboratory diagnosis of infections with Leishmania spp.


Assuntos
Leishmania , Leishmaniose , Psychodidae , Animais , Simulação por Computador , DNA de Protozoário , Leishmania/genética , Leishmaniose/diagnóstico , Leishmaniose/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária
17.
Acta Trop ; 217: 105851, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33524382

RESUMO

Schistosomiasis is a severe chronic disease caused by parasitic worms of the genus Schistosoma. Recent studies indicate that schistosomes can secrete extracellular vesicles (EVs), which play important regulatory roles in many biological processes. However, the mechanisms underlying EV biogenesis in schistosomes are poorly understood. In this study, we performed bioinformatic analyses and identified several genes putatively involved in EV biogenesis in Schistosoma japonicum, which were then confirmed by PCR. Quantitative transcriptional profiles of the selected genes indicated that they were differentially expressed in male and female worms as well as in the different developmental stages of S. japonicum. Thus, the highest expression of VAMP3 was detected in cercariae, whereas that of ARF6 was detected in eggs. RAB11A and the Syntenin-encoding gene SDCBP were highly expressed in 14-day schistosomula and VPS4A and RAB27A were highly expressed in 35-day-old adult schistosomes. The expression of RAB11A, CHMP4C, VPS4A, and SDCBP was higher in male worms, whereas that of ARF6, VAMP3, and RAB27A was higher in female worms. Our results are expected to provide important clues for understanding the role of EV biogenesis in S. japonicum development.


Assuntos
Vesículas Extracelulares/genética , Biogênese de Organelas , Schistosoma japonicum/crescimento & desenvolvimento , Schistosoma japonicum/genética , Esquistossomose/parasitologia , Transcriptoma , Animais , Fenômenos Biológicos , DNA de Protozoário , Vesículas Extracelulares/metabolismo , Feminino , Humanos , Estágios do Ciclo de Vida/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Óvulo/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
18.
Acta Trop ; 217: 105850, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33524385

RESUMO

Chronic opisthorchiasis caused by Opisthorchis viverrini (O. viverrini) adversely affects human health and is associated with a fatal bile duct cancer (cholangiocarcinoma). Although cats and dogs are known animal reservoir hosts of opisthorchiasis, there is limited information about whether other fish-eating animals are fluke reservoirs. Wetlands along Chi River of Thailand have abundant intermediate host snails and fish for O. viverrini and diverse other animal species. This led to our investigation into whether other fish-eating animals can also become infected and be alternate reservoir hosts for human opisthorchiasis. Our preliminary study focused on the O. viverrini infection status of crab-eating or long-tailed macaques roaming in Kosumpi National Forest Park in Chi River Basin, Kosumpisai District of Mahasarakam Province, and rodents, small residential mammals and fish-eating birds living in Lawa wetland complex of Khon Kaen Province. Fecal samples of each animal were collected and modified formalin ether concentration technique was applied to identify infections. Additionally, participatory epidemiology was used to retrieve information from local communities on opisthorchiasis transmission in these animals. No O. viverrini infection was found in any fecal samples suggesting that monkeys, rodents, small residential mammals and birds in these two wetlands probably do not serve as alternate reservoir hosts of O. viverrini.


Assuntos
Aves/parasitologia , Macaca/parasitologia , Opistorquíase/epidemiologia , Opisthorchis , Roedores/parasitologia , Animais , Gatos/parasitologia , Pesquisa Participativa Baseada na Comunidade , DNA de Protozoário , Reservatórios de Doenças/parasitologia , Cães/parasitologia , Estudos Epidemiológicos , Fezes/parasitologia , Humanos , Fígado/parasitologia , Óvulo/classificação , Contagem de Ovos de Parasitas , Reação em Cadeia da Polimerase , Prevalência , Tailândia/epidemiologia
19.
Acta Trop ; 217: 105858, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33582143

RESUMO

Chagas disease is an anthropozoonosis, caused by a flagellated protozoan, Trypanosoma cruzi, in which the enzootic cycle occurs between mammals and triatomines. Two dogs with a history of sudden death were necropsied at the Federal University of Pará (UFPA). One dog had a pale area in the myocardium, which on histopathological examination showed a T. cruzi amastigote nest; immunohistochemistry (IHC) analysis characterized it as acute Chagas disease (ACD). The second dog showed no macroscopic changes. Microscopically, a few cardiomyocytes were replaced by adipocytes, and IHC result was negative for T. cruzi. However, results of polymerase chain reaction (PCR) of the cardiac tissue of both dogs was positive for T. cruzi DNA. After that, an epidemiological study was conducted in the region. For this study, we selected four areas in Castanhal. One of the four areas (Area 1) is where one of the dogs lived. The other three areas were chosen because they were recently deforested for housing. Blood samples were collected from dogs, cats, wild small mammals (marsupials and rodents), and the digestive tract of triatomines. Nested PCR was performed on all the blood samples and the triatomine digestive tracts. In Area 1, T. cruzi DNA was detected in 50% (12/24) of the tested dogs, in the only tested cat (1/1), 50% (1/2) of the tested marsupials (Didelphis marsupials), and 100% of the captured triatomines (Rhodnius pictipes) (2/2). In Area 2, T. cruzi DNA was not detected in any of the 11 (0/11) dogs and two marsupials tested (0/2), and no triatomines were found in this area. In Area 3, T. cruzi DNA was detected in 42.25% (30/71) of the dogs, in 66,6% (2/3) of the cats, the only captured marsupial (D. marsupialis) (1/1), and all three triatomines (3/3) (R. pictipes) tested. In Area 4, the two dogs tested were negative (0/2), 25% (1/4) of the captured marsupials (D. marsupialis) was positive, and no triatomine was captured in this area. The data demonstrate the importance of detecting T. cruzi in dogs, cats, small rodents, and marsupials in the Amazon metropolitan areas, where ecotopes carry reservoirs and vectors capable of participating in the Chagas disease cycle. The proximity between humans and T. cruzi vectors in these places might contribute to increased disease transmission risk and maintenance of agents. It was noted that high-standard condominiums, previously thought to reduce the risk for this disease, presented a new epidemiological risk. The presence of T. cruzi DNA in a dog who, a year earlier had tested negative, when another dog in the same house died of ACD, shows that the transmission cycle is present and active, with a high possibility of disease transmission to animals and humans.


Assuntos
Doença de Chagas/veterinária , Doenças do Cão/parasitologia , Trypanosoma cruzi/genética , Animais , Doenças do Gato/parasitologia , Gatos/parasitologia , Doença de Chagas/transmissão , DNA de Protozoário , Didelphis/parasitologia , Doenças do Cão/diagnóstico , Doenças do Cão/patologia , Cães/parasitologia , Insetos Vetores/parasitologia , Mamíferos/parasitologia , Marsupiais/parasitologia , Reação em Cadeia da Polimerase , Rhodnius/parasitologia , Fatores de Risco , Roedores/parasitologia
20.
Rev Bras Parasitol Vet ; 30(1): e028520, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33605391

RESUMO

This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.


Assuntos
Doenças das Aves , Coccidiose , Sarcocystidae , Animais , Bioensaio , Doenças das Aves/epidemiologia , Doenças das Aves/parasitologia , Aves , Brasil , Coccidiose/epidemiologia , DNA de Protozoário , Camundongos , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Sarcocystidae/genética , Sarcocystis/genética , Toxoplasma/genética , Toxoplasmose Animal/epidemiologia
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